iBind Western Device

The original notebook-sized automated western blot processing device

Simply load your primary antibody, secondary antibody, and wash solutions into the device and then walk away. The Invitrogen iBind Western System automatically performs all immunoblotting steps using sequential lateral flow technology, a simple form of capillary action. In less than 3 hours, the blot is ready for final detection. You can continue to use your existing chromogenic, chemiluminescent, or fluorescent western blot detection protocols, along with your choice of primary antibody or secondary antibody conjugates of HRP, AP, or fluorescent dyes.

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Example data

Robust results with less primary antibody

One of the key elements of a successful western blot is the primary antibody; however, this reagent also contributes to over 90% of the total cost of the blot. Because the iBind Western System is more sensitive than manual processing methods, you can use less antibody and achieve similar results.

*Results may vary. The protocol for primary antibody use is 80% less than a traditional manual method.

Using chemiluminescent or fluorescence-based detection methods, you can use up to 80% less primary antibody than manual methods, saving you significant cost per blot. A 2-fold dilution series of EGF receptor control cell lysate (30 µg, 15 µg, 7.5 µg, 3.75 µg, and 1.875 µg) was used. Proteins were separated using Bolt Bis-Tris Plus gels on the Mini Gel Tank and transferred to PVDF membranes using the iBlot 2 Dry Blotting System. The blots were probed with an anti–phospho-EGF receptor [Tyr1068] (1H12) mouse monoclonal antibody (1:1,000 dilution, equated to 2 µL antibody for the iBind device method and 10 µL antibody for the manual method) followed by a goat anti–mouse IgG (H+L) peroxidase-conjugated antibody (1:360 for iBind device processing (5.55 µL); 1:1,800 for manual method (5.55 µL)). The standard iBind solution kit was used for the chemiluminescence blot (panel C, manual processing and panel D, iBind system processing); the fluorescence detection (panel A, manual processing and panel B, iBind processing) kit was used for fluorescence detection. These results demonstrate that western blots processed on the iBind device show comparable results to those obtained when western blots are processed manually, with lower overall primary antibody requirements for the iBind device.

Excellent western performance compared to manual blotting

The iBind System offers greater sensitivity compared to manual methods for many monoclonal and polyclonal antibodies.

Combine the iBind Western System with highly specific primary and secondary antibodies to achieve cleaner western blots.

Automated processing enables improved blot-to-blot consistency with CVs typically less than 5% (compared to manual processing that can have CVs of 13%).

Western blots processed on the Invitrogen iBind device show excellent sensitivity compared to western blots processed manually. (A–B) Western blots with phosphorylated Akt (left to right: 30 µg–500 ng cell lysate load) were processed either on the iBind device or using standard manual western processing protocols as specified by the antibody manufacturer (monoclonal anti–phospho-Akt [pT308] (C31E5E) primary antibody; HRP-conjugated anti-rabbit secondary antibody). The blot processed with the iBind device detected phospho-Akt in 500 ng of cell lysate, while the target was detected in 4 µg on the manually processed blot. (C–D) Western blots with cell lysate expressing CREB (left to right: 30 µg–1 µg cell lysate load) were processed either on the iBind device or using standard manual western processing protocols as specified by the antibody manufacturer (polyclonal anti-CREB primary antibody; HRP-conjugated anti-rabbit secondary antibody). The blot processed with the iBind system detected CREB in 6 µg of cell lysate, while 10 µg of lysate was needed to detect CREB on the manually processed blot. For all blots, proteins were separated using Bolt Bis-Tris Plus gels on the Mini Gel Tank and transferred to PVDF membranes using the iBlot 2 Dry Blotting System.

Robust results with less primary antibody

One of the key elements of a successful western blot is the primary antibody; however, this reagent also contributes to over 90% of the total cost of the blot. Because the iBind Western System is more sensitive than manual processing methods, you can use less antibody and achieve similar results.

*Results may vary. The protocol for primary antibody use is 80% less than a traditional manual method.

Using chemiluminescent or fluorescence-based detection methods, you can use up to 80% less primary antibody than manual methods, saving you significant cost per blot. A 2-fold dilution series of EGF receptor control cell lysate (30 µg, 15 µg, 7.5 µg, 3.75 µg, and 1.875 µg) was used. Proteins were separated using Bolt Bis-Tris Plus gels on the Mini Gel Tank and transferred to PVDF membranes using the iBlot 2 Dry Blotting System. The blots were probed with an anti–phospho-EGF receptor [Tyr1068] (1H12) mouse monoclonal antibody (1:1,000 dilution, equated to 2 µL antibody for the iBind device method and 10 µL antibody for the manual method) followed by a goat anti–mouse IgG (H+L) peroxidase-conjugated antibody (1:360 for iBind device processing (5.55 µL); 1:1,800 for manual method (5.55 µL)). The standard iBind solution kit was used for the chemiluminescence blot (panel C, manual processing and panel D, iBind system processing); the fluorescence detection (panel A, manual processing and panel B, iBind processing) kit was used for fluorescence detection. These results demonstrate that western blots processed on the iBind device show comparable results to those obtained when western blots are processed manually, with lower overall primary antibody requirements for the iBind device.

The iBind System offers greater sensitivity compared to manual methods for many monoclonal and polyclonal antibodies.

Combine the iBind Western System with highly specific primary and secondary antibodies to achieve cleaner western blots.

Automated processing enables improved blot-to-blot consistency with CVs typically less than 5% (compared to manual processing that can have CVs of 13%).

Western blots processed on the Invitrogen iBind device show excellent sensitivity compared to western blots processed manually. (A–B) Western blots with phosphorylated Akt (left to right: 30 µg–500 ng cell lysate load) were processed either on the iBind device or using standard manual western processing protocols as specified by the antibody manufacturer (monoclonal anti–phospho-Akt [pT308] (C31E5E) primary antibody; HRP-conjugated anti-rabbit secondary antibody). The blot processed with the iBind device detected phospho-Akt in 500 ng of cell lysate, while the target was detected in 4 µg on the manually processed blot. (C–D) Western blots with cell lysate expressing CREB (left to right: 30 µg–1 µg cell lysate load) were processed either on the iBind device or using standard manual western processing protocols as specified by the antibody manufacturer (polyclonal anti-CREB primary antibody; HRP-conjugated anti-rabbit secondary antibody). The blot processed with the iBind system detected CREB in 6 µg of cell lysate, while 10 µg of lysate was needed to detect CREB on the manually processed blot. For all blots, proteins were separated using Bolt Bis-Tris Plus gels on the Mini Gel Tank and transferred to PVDF membranes using the iBlot 2 Dry Blotting System.